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Very high activity biocatalysts for low water systems: Propanol rinsed enzyme preparations (PREPs)

Moore, B.D. and Partridge, J. and Halling, P.J. (2001) Very high activity biocatalysts for low water systems: Propanol rinsed enzyme preparations (PREPs). In: Methods in Biotechnology: Enzymes in Nonaqueous Solvents. Methods in Biotechnology, 15 . Humana Press, NJ, USA, pp. 97-104. ISBN 0896039293

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Abstract

Depending on the preparation method used enzymes under low water conditions can exhibit differences in catalytic activity that vary by several orders of magnitude. Since this can make the difference between whether a particular biotransformation is practically useful or not it is important to control enzyme history carefully. The basic problem is simple: how to transfer the enzyme from an aqueous environment to one where it is dehydrated whilst ensuring firstly it remains in a native conformation and secondly the active site residues are in the correct protonation state. Although the importance of retaining native structure is well recognised potential detrimental changes to protonation state are often not considered. Usually it is assumed that the enzyme will exhibit 'pH memory' of the previous aqueous solution. As discussed in detail in chapter ?? Partidge this is not always a valid assumption.